PCRmax Yersinia enterocolitica Instruction Manual

Attachment invasion locus protein
(ail) gene

Yersinia enterocolitica

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Yersinia enterocolitica belongs to a family of rod-shaped bacteria termed
Enterobacteriaceae. They consist of 11 species, three of which are pathogenic to humans:
Yersinia pestis and the enteropathogenic yersiniae; Yersinia pseudotuberculosis and
Yersinia enterocolitica. All three species target the lymph tissues during infection and carry
a 70 kb virulence plasmid (pYV), which is essential for infection in these tissues as well as
to overcome host defence mechanisms. Our kit specifically targets the AIL gene
(Attachment invasion locus protein) of Y. enterocolitica.

The major animal reservoir for Y. enterocolitica strains that cause human illness is pigs,
but other strains are also found in many other animals including rodents, rabbits, sheep,
cattle, horses, dogs, and cats. In pigs, the bacteria are most likely to be found on the
tonsils.

Infection is most often acquired by eating contaminated food, especially raw or
undercooked pork products. The preparation of raw pork intestines may be particularly
riskyDrinking contaminated unpasteurized milk or untreated water can also transmit the
infection. Occasionally Y. enterocoliticainfection occurs after contact with infected animals.
On rare occasions, it can be transmitted as a result of the bacterium passing from the
stools or soiled fingers of one person to the mouth of another person. This may happen
when basic hygiene and handwashing habits are inadequate. Rarely, the organism is
transmitted through contaminated blood during a transfusion. Children are infected more
often than adults, and the infection is more common in the winter.

Yersinia enterocolitica is a psychotropic bacterium, which causes acute gastro-enteritis
and occasionally more serious disease in humans. In some countries it rivals Salmonella
as a food borne pathogen and because it can grow at refrigeration temperature it is an
increasing concern in terms of food safety. Acute Y. enterocolitica infections usually lead to
mild self-limiting entero-colitis or terminal ileitis in humans. Symptoms may include watery
or bloody diarrhea and fever. After oral uptake yersiniae replicate in the terminal ileum and
invade Peyer's patches. From here yersiniae can disseminate further to mesenteric lymph
nodes causing lymphadenopathy. This condition can be confused with appendicitis and is
therefore called pseudoappendicitis.

In immunosuppressed individuals, yersiniae can disseminate from the gut to liver and
spleen and form abscesses. Because Yersinia is a siderophilic (iron-loving) bacteria,
people with hereditary hemochromatosis (a disease resulting in high body iron levels) are
more susceptible to infection.

Introduction to Yersinia enterocolitica

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MIN

MAX

The PCR Max qPCR Kit for Yersinia enterocolitica (Y.enterocolitica) genomes is designed
for the in vitro quantification of Y.enterocolitica genomes. The kit is designed to have the
broadest detection profile possible whilst remaining specific to the Y.enterocolitica
genome.

The primers and probe sequences in this kit have 100% homology with a broad range of
Y.enterocolitica sequences based on a comprehensive bioinformatics analysis.

If you require further information, or have a specific question about the detection profile of
this kit then please send an e.mail to [e-mail] and our bioinformatics team will
answer your question.

Specificity

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Kit Contents

• Y.enterocolitica specific primer/probe mix (150 reactions BROWN)

FAM labelled

• Y.enterocolitica positive control template (for Standard curve RED)

• Internal extraction control primer/probe mix (150 reactions BROWN)

VIC labelled as standard

• Internal extraction control DNA (150 reactions BLUE)

• Endogenous control primer/probe mix (150 reactions BROWN)

FAM labelled

• RNAse/DNAse free water (WHITE)

for resuspension of primer/probe mixes and internal extraction control DNA

• Template preparation buffer (YELLOW)

for resuspension of positive control template and standard curve preparation

Reagents and equipment to be supplied by the user

Real-Time PCR Instrument

DNA extraction kit

This kit designed to work well with all processes that yield high quality DNA with minimal
PCR inhibitors.

Lyophilised 2x qPCR Mastermix

This kit is designed to be compatible with all commercially available Mastermixes that run
with standard cycling conditions.

Pipettors and Tips

Vortex and centrifuge

Thin walled 1.5 ml PCR reaction tubes

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